Department of Urology, Shanghai Tenth People's Hospital, Tongji University, Shanghai 200072, China.
Department of Urology, The Affiliated People's Hospital of Ningbo University, 251 East Baizhang Road, Ningbo City, Zhejiang Province, China.
Biomed Res Int. 2020 Nov 17;2020:4817608. doi: 10.1155/2020/4817608. eCollection 2020.
The long noncoding RNA LUCAT1 (lung cancer-associated transcript 1) has been reported to be highly expressed in bladder cancer samples, but its role and molecular mechanisms need to be elucidated.
Bioinformatics methods show that miR-181c-5p is a target of LUCAT1. Here, we aimed to reveal whether LUCAT1 participates in the development of bladder cancer via targeting miR-181c-5p. The expression levels of LUCAT1 and miR-181c-5p were detected by RT-PCR technology in bladder cells and tissues. The effects of the LUCAT1/miR-181c-5p axis on cell proliferation, migration, invasion, and apoptosis were tested by CCK-8, wound healing, Transwell chambers, and flow cytometry assays. The expressions of apoptosis/migration-related proteins were detected by western blotting assays.
The results demonstrated that LUCAT1 was overexpressed in bladder cancer tissue and cells, while miR-181c-5p showed a low expression pattern as compared to normal bladder cells and tissues. Cell proliferation, migration, and invasion capacities were significantly impaired, and cell apoptosis was enhanced when LUCAT1 was silenced in UM-UC-3 and T24 cell lines, but this effect was abolished by miR-181c-5p downregulation. In addition, miR-181c-5p downregulation impaired LUCAT1 downregulation which mediated the decreased expressions of Bcl2 and N-cadherin and the increased expressions of Bax and E-cadherin. Moreover, we found that KRAS was a direct target of miR-181c-5p and was under the positive regulation of LUCAT1.
Collectively, this study reveals that knockdown of LUCAT1 inhibits the migration and invasion of bladder cancer cells in a miR-181c-5p-dependent manner, which may be related to KRAS downregulation.
长链非编码 RNA LUCAT1（肺癌相关转录物 1）已被报道在膀胱癌样本中高度表达，但它的作用和分子机制仍需阐明。
生物信息学方法表明 miR-181c-5p 是 LUCAT1 的一个靶标。在这里，我们旨在通过靶向 miR-181c-5p 来揭示 LUCAT1 是否参与膀胱癌的发生。通过 RT-PCR 技术检测膀胱细胞和组织中 LUCAT1 和 miR-181c-5p 的表达水平。通过 CCK-8、划痕愈合、Transwell 室和流式细胞术检测 LUCAT1/miR-181c-5p 轴对细胞增殖、迁移、侵袭和凋亡的影响。通过 Western blot 检测凋亡/迁移相关蛋白的表达。
结果表明，LUCAT1 在膀胱癌组织和细胞中过度表达，而 miR-181c-5p 的表达模式与正常膀胱细胞和组织相比较低。当 UM-UC-3 和 T24 细胞系中沉默 LUCAT1 时，细胞增殖、迁移和侵袭能力显著受损，细胞凋亡增强，但这种作用被 miR-181c-5p 下调所消除。此外，miR-181c-5p 下调削弱了 LUCAT1 下调，导致 Bcl2 和 N-cadherin 的表达减少，Bax 和 E-cadherin 的表达增加。此外，我们发现 KRAS 是 miR-181c-5p 的直接靶标，并且受 LUCAT1 的正向调节。
总之，本研究表明，沉默 LUCAT1 以 miR-181c-5p 依赖的方式抑制膀胱癌细胞的迁移和侵袭，这可能与 KRAS 下调有关。
